Diene Compounds and Formulations

ABSTRACT

The instant invention provides potent antiandrogen compounds, such as 3β-acetoxyandrost-1,5-diene-17-ethylene ketal and 3β-hydroxyandrost-1,5-diene-17-ethylene ketal, and methods for their use in the prevention and treatment of biological conditions mediated by androgen receptors. Thus, for example, compounds of the invention are useful in the prevention and treatment of prostrate cancer. Furthermore, it has been discovered that compounds of the invention are useful in the prevention and treatment of androgen-independent cancers such as androgen-independent prostrate cancer. Finally, inventive compounds may be used to treat antiandrogen induced withdrawal syndrome.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority from and is a continuation applicationof pending application Ser. No. 11/838,154, filed Aug. 13, 2007, whichis a continuation application of abandoned U.S. application Ser. No.10/814,503, filed Mar. 30, 2004, which claims priority from abandonedU.S. provisional application No. 60/459,450, filed on Apr. 1, 2003, allthree of which applications are incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to low molecular weight compounds, usefulas antiandrogens, and in particular as antiandrogens with low androgenicactivity. The invention further relates to methods for inhibitingandrogen receptors and in treating androgen receptor-mediatedconditions, such as prostate cancer, with compounds and compositionsprovided herein.

BACKGROUND OF THE INVENTION

Androgens play a major role in promoting the development and progressionof prostate cancer. Consequently, since the first observation by Hugginsand Hodges in 1941 (Cancer Res., 1941, 1:293-297), endocrine therapyremains the critical therapeutic option for advanced forms of prostatecancer. This therapy consists of androgen ablation by medical orsurgical castration and/or inhibiting the receptor level action ofandrogens from both the testes and adrenal glands by antiandrogens.Thus, antiandrogens are generally used in conjunction with castration ascombined androgen blockade (CAB). Unfortunately, after a brief clinicalresponse to the hormonal therapy in most patients, the majorityeventually develop symptomatic recurrences, which have been termedandrogen-independent or hormone-refractory prostate cancer, within a fewyears. Indeed, in males prostate cancer is the most common malignancyand is the second leading cause of cancer-related death.

Antiandrogens include a number of compounds that are able to competewith androgens, such as dihydrotestosterone (DHT), an active metaboliteof testosterone in the prostate, for the binding to the androgenreceptor (AR). There are three non-steroidal antiandrogens available inthe United States: flutamide, bicalutamide (casodex), and nilutamide.Monotherapy using these antiandrogens does not decrease androgenconcentrations, offering potential quality-of-life benefits overcastration-based approaches. However, specific side effects may beassociated with such monotherapy, including gynecomastia and breastpain, hepatotoxicity, visual and respiratory disturbances, and alcoholintolerance (Kolvenbag, et al., Urology, 2001, 58(Suppl 2A):16-23.).

In addition, antiandrogens have been reported to raise the amount ofprostate-specific antigen (PSA), a tumor marker of prostate cancer andalso an AR responsive gene, during hormonal therapy. In these cases,when antiandrogen therapy is terminated, PSA actually declines to 50% orless of its original value prior to therapy; this phenomenon is known asantiandrogen withdrawal syndrome. Thus, such patients benefit from thewithdrawal of the majority of antiandrogens clinically used, includingthe above three drugs, as well as some steroid hormones, such asdiethlystilbesterol and magestrol. The mechanisms responsible forantiandrogen withdrawal syndrome are not completely understood, althoughit is likely that AR gene mutations and/or AR coregulators, such asARA70, are involved in the change of antiandrogens from antagonists toagonists. The remaining patients, not subject to antiandrogen withdrawalsyndrome may be considered to have androgen-independent prostate cancer.

Thus, a need in the art exists for new and more effective antiandrogeniccompounds with lower androgenic activities. In particular, there is aneed for antiandrogenic compounds effective against prostate cancer andespecially against androgen-independent prostate cancer.

Methods to analyze or characterize the effects of androgen receptormodulators, i.e., agonists and antagonists, have been described, e.g.,Yeh, S., et al. (1997) Lancet 349, 852-853; Miyamoto, H., et al. (1998)Proc. Natl. Acad. Sci. USA 95, 7379-7384; Miyamoto, H., et al. (1998)Proc. Natl. Acad. Sci. USA 95, 11083-11088; Chang, H.-C., et al. (1999)Proc. Natl. Acad. Sci. USA 96, 11173-11177; Miyamoto, H., et al. (2003)Proc. Natl. Acad. Sci. USA 100, 4440-4444; Rahman, M. M., et al. (2003)Proc. Natl. Acad. Sci. USA 100, 5124-5129; Yeh, S. & Chang, C. (1996)Proc. Natl. Acad. Sci. USA 93, 5517-5521.

SUMMARY OF THE INVENTION

The instant invention provides potent antiandrogen compounds and methodsfor their use in the prevention and treatment of biological conditionsmediated by androgen receptors. Thus, for example, compounds of theinvention are useful in the prevention and treatment of prostate cancer.Furthermore, it has been discovered that compounds of the invention areuseful in the prevention and treatment of androgen-independent cancerssuch as androgen-independent prostrate cancer. Finally, inventivecompounds may be used to treat antiandrogen induced withdrawal syndrome.

Thus, there has been provided, in accordance with one aspect of theinvention, compounds of structure I, prodrugs thereof, pharmaceuticallyacceptable salts thereof, stereoisomers thereof, tautomers thereof, andsolvates thereof.

In compounds having structure I

A is —C(O)—, ═CR⁹—, or —CR⁹R¹⁰—;

E is —C(O)—, ═CR⁵—, or —CR⁵R⁶—, wherein A and E are not both —C(O);

G is —C(O)—, ═CR³—, or —CR³R⁴—;

K is —C(O)—, ═CR¹—, or —CR¹R²—;

R¹ is selected from the group consisting of —OR¹¹, substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, substituted and unsubstituted heterocyclylalkyl,—C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹², —C(S)—OR¹², —NR¹²R¹³,—NR¹²—C(O)—R¹³, —NR¹²—C(O)—OR¹³, —NR¹²—C(O)—NR¹²R¹³, and —S(O)₀₋₂—R¹²;

R² is selected from the group consisting of —H, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,and substituted and unsubstituted lower alkyne;

R³ and R⁵ are independently selected from the group consisting of —H,—F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl, substitutedand unsubstituted lower alkene, substituted and unsubstituted loweralkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH,substituted and unsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴;

R⁴ and R⁶ are independently selected from the group consisting of —H,—F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl, substitutedand unsubstituted lower alkene, substituted and unsubstituted loweralkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH,substituted and unsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴;

R⁷ and R⁸ are independently selected from the group consisting of —H andsubstituted and unsubstituted lower alkyl group;

R⁹ and R¹⁰ are independently selected from the group consisting ofsubstituted and unsubstituted lower alkyl, substituted and unsubstitutedlower alkene, substituted and unsubstituted lower alkyne, —OH, whereinR⁹ and R¹⁰ are not both —OH, substituted and unsubstituted lower alkoxy,and substituted and unsubstituted —S(O)₀₋₂(lower alkyl), or R⁹ and R¹⁰,together with the carbon to which they are attached, form a 5-, 6-, or7-membereterocyclyl or cycloalkyl group;

R¹¹ is selected from the group consisting of —H, substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, substituted and unsubstituted heterocyclylalkyl,—C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹², —NR¹²R¹³, —S(O)₂—R¹²,—S(O)₂—OR¹², or —P(O)(OR¹²)(OR¹³)₀₋₁;

R¹² and R¹³ are, at each occurrence, independently selected from thegroup consisting of —H, substituted and unsubstituted alkyl, substitutedand unsubstituted alkene, substituted and unsubstituted alkyne,substituted and unsubstituted aryl, substituted and unsubstitutedarylalkyl, substituted and unsubstituted heterocyclyl, and substitutedand unsubstituted heterocyclylalkyl;

R¹⁴ and R¹⁵ are, at each occurrence, independently selected from thegroup consisting of substituted and unsubstituted lower alkyl,substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, substituted and unsubstituted C₆₋₁₀ aryl,and substituted and unsubstituted C₇₋₁₂ arylalkyl;

R and R′ are, at each occurrence, independently selected from the groupconsisting of —F, —Cl, —Br, —I, substituted and unsubstituted loweralkyl, substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted lower alkoxy, and—S(O)₀₋₂R¹⁴;

n and n′ are independently 0, 1, or 2; and

wherein the dashed lines in structure I represent carbon-carbon doublebonds or carbon-carbon single bonds contained within the fused four-ringsystem, such that the compound comprises a 1,6-diene, 1,7-diene,1,8-diene, 1,15-diene, 1,16-diene, 4,8-diene, 3,16-diene, 1,3,5-triene,1,3,16-triene, 1,5,7-triene, 1,5,15-triene, 1,8,15-triene,1,5,16-triene, or 1,5,7,15-tetraene, within the fused four-ring system.

In another aspect of the invention, there are provided compounds ofstructure II, prodrugs thereof, pharmaceutically acceptable saltsthereof, stereoisomers thereof, tautomers thereof, or solvates thereof.

In compounds of structure II,

A is —C(O)— or —CR⁹R¹⁰—,

E is —C(O)— or —CR⁵R⁶—, wherein A and E are not both —C(O)—;

G is —C(O)—, ═CR³—, or —CR³R⁴—;

K is ═C(OR¹¹)—, or —C(OR¹¹)R²—;

R² is selected from the group consisting of —H, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,and substituted and unsubstituted lower alkyne;

R³ and R⁵ are independently selected from the group consisting of —H,—F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl, substitutedand unsubstituted lower alkene, substituted and unsubstituted loweralkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH,substituted and unsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴;

R⁴ and R⁶ are independently selected from the group consisting of —H,—F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl, substitutedand unsubstituted lower alkene, substituted and unsubstituted loweralkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH,substituted and unsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴;

R⁷ and R⁸ are independently selected from the group consisting of —H andsubstituted and unsubstituted lower alkyl group;

R⁹ and R¹⁰ are independently selected from the group consisting ofsubstituted and unsubstituted lower alkyl, substituted and unsubstitutedlower alkene, substituted and unsubstituted lower alkyne, —OH, whereinR⁹ and R¹⁰ are not both —OH, substituted and unsubstituted lower alkoxy,and substituted and unsubstituted —S(O)₀₋₂(lower alkyl), or R⁹ and R¹⁰,together with the carbon to which they are attached, form a 5-, 6-, or7-member heterocyclyl or cycloalkyl group;

R¹¹ is selected from the group consisting of substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, substituted and unsubstituted heterocyclylalkyl,—C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹², —NR¹²R¹³, —S(O)₂—R¹²,—S(O)₂—OR¹² or —P(O)(OR¹²)(OR¹³)₀₋₁;

R¹² and R¹³ are, at each occurrence, independently selected from thegroup consisting of —H, substituted and unsubstituted alkyl, substitutedand unsubstituted alkene, substituted and unsubstituted alkyne,substituted and unsubstituted aryl, substituted and unsubstitutedarylalkyl, substituted and unsubstituted heterocyclyl, and substitutedand unsubstituted heterocyclylalkyl;

R¹⁴ and R¹⁵ are, at each occurrence, independently selected from thegroup consisting of substituted and unsubstituted lower alkyl,substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, substituted and unsubstituted C₆₋₁₀ aryl,and substituted and unsubstituted C₇₋₁₂ arylalkyl;

R and R′ are, at each occurrence, independently selected from the groupconsisting of —F, —Cl, —Br, —I, substituted and unsubstituted loweralkyl, substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted lower alkoxy, and—S(O)₀₋₂R¹⁴;

n and n′ are independently 0, 1, or 2; and

wherein the dashed lines in structure II represent carbon-carbon doublebonds or carbon-carbon single bonds contained within the fused four-ringsystem, such that the compound comprises a 1,3-diene, 1,5-diene, or1,4,6-triene within the fused four-ring system.

In still another aspect of the invention, there are provided methods oftreating or preventing a condition mediated by an androgen receptorcomprising administering to a subject in need thereof, an effectiveamount of a compound having the structure III, prodrugs thereof,pharmaceutically acceptable salts thereof, stereoisomers thereof,tautomers thereof, or solvates thereof.

In compounds of structure III

A is —C(O)—, ═CR⁹—, or —CR⁹R¹⁰—;

E is —C(O)—, ═CR⁵—, or —CR⁵R⁶—, wherein A and E are not both —C(O);

G is —C(O)—, ═CR³—, or —CR³R⁴—;

K is —C(O)—, ═CR¹—, or —CR¹R²—;

R¹ is selected from the group consisting of —OR¹¹, substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, substituted and unsubstituted heterocyclylalkyl,—O—C(O)—R¹², —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹²,—C(S)—OR¹², —NR¹²R¹³, —NR¹²—C(O)—R¹³, —NR¹²—C(O)—OR¹³,—NR¹²—C(O)—NR¹²R¹³, and —S(O)₀₋₂—R²;

R² is selected from the group consisting of —H, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,and substituted and unsubstituted lower alkyne;

R³ and R⁵ are independently selected from the group consisting of —H,—F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl, substitutedand unsubstituted lower alkene, substituted and unsubstituted loweralkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH,substituted and unsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴;

R⁴ and R⁶ are independently selected from the group consisting of —H,—F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl, substitutedand unsubstituted lower alkene, substituted and unsubstituted loweralkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH,substituted and unsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴;

R⁷ and R⁸ are independently selected from the group consisting of —H andsubstituted and unsubstituted lower alkyl group;

R⁹ and R¹⁰ are independently selected from the group consisting ofsubstituted and unsubstituted lower alkyl, substituted and unsubstitutedlower alkene, substituted and unsubstituted lower alkyne, —OH, whereinR⁹ and R¹⁰ are not both —OH, substituted and unsubstituted lower alkoxy,and substituted and unsubstituted —S(O)₀₋₂(lower alkyl), or R⁹ and R¹⁰,together with the carbon to which they are attached, form a 5-, 6-, or7-member heterocyclyl or cycloalkyl group;

R¹¹ is selected from the group consisting of —H, substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, substituted and unsubstituted heterocyclylalkyl,—C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹², —NR¹²R¹³, —S(O)R¹²,—S(O)₂—R¹² or —P(O)(OR¹²)(OR¹³)₀₋₁;

R¹² and R¹³ are, at each occurrence, independently selected from thegroup consisting of —H, substituted and unsubstituted alkyl, substitutedand unsubstituted alkene, substituted and unsubstituted alkyne,substituted and unsubstituted aryl, substituted and unsubstitutedarylalkyl, substituted and unsubstituted heterocyclyl, and substitutedand unsubstituted heterocyclylalkyl;

R¹⁴ and R¹⁵ are, at each occurrence, independently selected from thegroup consisting of substituted and unsubstituted lower alkyl,substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, substituted and unsubstituted C₆₋₁₀ aryl,and substituted and unsubstituted C₇₋₁₂ arylalkyl;

R and R′ are, at each occurrence, independently selected from the groupconsisting of —F, —Cl, —Br, —I, substituted and unsubstituted loweralkyl, substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted lower alkoxy, and—S(O)₀₋₂R¹⁴;

n and n′ are independently 0, 1, or 2; and

wherein the dashed lines in structure III represent carbon-carbon doublebonds or carbon-carbon single bonds contained within the fused four-ringsystem, such that the compound comprises a 1,3-diene, 1,5-diene,1,6-diene, 1,7-diene, 1,8-diene, 1,15-diene, 1,16-diene, 3,16-diene,4,8-diene, 1,3,5-triene, 1,4,6-triene, 1,3,16-triene, 1,5,7-triene,1,5,15-triene, 1,8,15-triene, 1,5,16-triene, or 1,5,7,15-tetraene,within the fused four-ring system.

In some embodiments of methods of treating or preventing a conditionmediated by an androgen receptor, the condition is prostate cancer, andin particular, prostate cancer at an androgen-independent stage. Inother embodiments, the condition is antiandrogen induced withdrawalsyndrome, and the subject may be afflicted with prostate cancer.

In other embodiments of methods of treating or preventing a conditionmediated by an androgen receptor, the compound comprises a 1,5-dienewithin the fused four-ring system. In other such embodiments, K is—CR¹R²—, R¹ is —OR¹¹, or R¹¹ is —H, substituted or unsubstituted alkyl,—C(O)—R¹², —C(O)—NR¹²R¹³, or —C(O)—OR¹². In further embodiments, thecompound comprises a 1,5-diene within the fused four-ring system, andR¹¹ is —H, or —C(O)—R¹².

The present invention also provides methods of inhibiting androgenreceptors in vitro or in vivo comprising contacting an androgen receptorwith an effective amount of a compound having the structure III, asdescribed above. In some embodiments of such methods, thetransactivation of androgen receptor is suppressed. In otherembodiments, the androgen receptor is mutant or native androgenreceptor.

In still other embodiments of methods of inhibiting androgen receptorsin vitro or in vivo, the compound comprises a 1,5-diene within the fusedfour-ring system. In other such embodiments, K is —CR¹R²—, R¹ is —OR¹¹,or R¹¹ is —H, substituted or unsubstituted alkyl, —C(O)—R¹²,—C(O)—NR¹²R¹³, or —C(O)—OR¹². In further embodiments, the compoundcomprises a 1,5-diene within the fused four-ring system, and R¹¹ is —H,or —C(O)—R¹².

DETAILED DESCRIPTION OF THE INVENTION

The following abbreviations and terms are defined as follows: ADEK is3β-acetoxy-17,17-ethylenedioxyandrost-1,5-diene-17-one(3β-acetoxyandrost-1,5-diene-17-ethylene ketal), adiol isΔ⁵-androstenediol (3β,17β-dihydroxyandrost-5-ene), AR is androgenreceptor, DHEA is dehydroepiandrosterone, DHT is dihydrotestosterone, ERis estrogen receptor, EtOH is ethanol, HF is hydroxyflutamide, Luc isluciferase, MMTV is mouse mammary tumor virus, PR is progesteronereceptor, PSA is prostrate specific antigen, R1881 is the syntheticandrogen methyltrienolone(17α-methyl-17β-hydroxyestra-4,9(10),11-trien-3-one) and RBA is relativebinding affinity.

Generally, reference to a certain element such as hydrogen or —H ismeant to include all isotopes of that element. For example, if an Rgroup is defined to include hydrogen or —H, it also includes deuteriumand tritium.

The phrase “unsubstituted alkyl” refers to alkyl groups that do notcontain heteroatoms. Thus the phrase includes straight chain alkylgroups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl,octyl, nonyl, decyl, undecyl, dodecyl and the like. The phrase alsoincludes branched chain isomers of straight chain alkyl groups,including but not limited to, the following which are provided by way ofexample: —CH(CH₃)₂, —CH(CH₃)(CH₂CH₃), —CH(CH₂CH₃)₂, —C(CH₃)₃,—C(CH₂CH₃)₃, —CH₂CH(CH₃)₂, —CH₂CH(CH₃)(CH₂CH₃), —CH₂CH(CH₂CH₃)₂,—CH₂C(CH₃)₃, —CH₂C(CH₂CH₃)₃, —CH(CH₃)CH(CH₃)(CH₂CH₃), —CH₂CH₂CH(CH₃)₂,—CH₂CH₂CH(CH₃)(CH₂CH₃), —CH₂CH₂CH(CH₂CH₃)₂, —CH₂CH₂C(CH₃)₃,—CH₂CH₂C(CH₂CH₃)₃, —CH(CH₃)CH₂CH(CH₃)₂, —CH(CH₃)CH(CH₃)CH(CH₃)₂,—CH(CH₂CH₃)CH(CH₃)CH(CH₃)(CH₂CH₃), and others. The phrase also includescyclic alkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl and such rings substituted withstraight and branched chain alkyl groups as defined above. The phrasealso includes polycyclic alkyl groups such as, but not limited to,adamantyl, norbornyl, and bicyclo[2.2.2]octyl and such rings substitutedwith straight and branched chain alkyl groups as defined above. Thus,the phrase unsubstituted alkyl groups includes primary alkyl groups,secondary alkyl groups, and tertiary alkyl groups. Unsubstituted alkylgroups may be bonded to one or more carbon atom(s), oxygen atom(s),nitrogen atom(s), and/or sulfur atom(s) in the parent compound.Preferred unsubstituted alkyl groups include straight and branched chainalkyl groups and cyclic alkyl groups having 1 to 20 carbon atoms, andmore preferred such groups have from 1 to 10 carbon atoms. Even morepreferred such groups, also known as unsubstituted lower alkyl groups,have from 1 to 5 carbon atoms. Most preferred unsubstituted alkyl groupsinclude straight and branched chain alkyl groups having from 1 to 3carbon atoms and include methyl, ethyl, propyl, and —CH(CH₃)₂.

The phrase “substituted alkyl” refers to an unsubstituted alkyl group asdefined above in which one or more bonds to a carbon(s) or hydrogen(s)are replaced by a bond to non-hydrogen and non-carbon atoms such as, butnot limited to, a halogen atom in halides such as F, Cl, Br, and I; andoxygen atom in groups such as hydroxyl groups, alkoxy groups, aryloxygroups, and ester groups; a sulfur atom in groups such as thiol groups,alkyl and aryl sulfide groups, sulfone groups, sulfonyl groups, andsulfoxide groups; a nitrogen atom in groups such as amines, amides,alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines,N-oxides, imides, and enamines; a silicon atom in groups such as intrialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups,and triarylsilyl groups; and other heteroatoms in various other groups.Substituted alkyl groups also include groups in which one or more bondsto a carbon(s) or hydrogen(s) atom is replaced by a bond to a heteroatomsuch as oxygen in carbonyl, carboxyl, and ester groups; nitrogen ingroups such as imines, oximes, hydrazones, and nitriles. Preferredsubstituted alkyl groups include, among others, alkyl groups in whichone or more bonds to a carbon or hydrogen atom is/are replaced by one ormore bonds to fluorine atoms. One example of a substituted alkyl groupis the trifluoromethyl group and other alkyl groups that contain thetrifluoromethyl group. Other alkyl groups include those in which one ormore bonds to a carbon or hydrogen atom is replaced by a bond to anoxygen atom such that the substituted alkyl group contains a hydroxyl,alkoxy, aryloxy group, or heterocyclyloxy group. Still other alkylgroups include alkyl groups that have an amine, alkylamine,dialkylamine, arylamine, (alkyl)(aryl)amine, diarylamine,heterocyclylamine, (alkyl)(heterocyclyl)amine,(aryl)(heterocyclyl)amine, or diheterocyclylamine group.

The phrase “unsubstituted aryl” refers to aryl groups that do notcontain heteroatoms. Thus the phrase includes, but is not limited to,groups such as phenyl, biphenyl, anthracenyl, naphthenyl by way ofexample. Although the phrase “unsubstituted aryl” includes groupscontaining condensed rings such as naphthalene, it does not include arylgroups that have other groups such as alkyl or halo groups bonded to oneof the ring members, as aryl groups such as tolyl are considered hereinto be substituted aryl groups as described below. A preferredunsubstituted aryl group is phenyl. Unsubstituted aryl groups may bebonded to one or more carbon atom(s), oxygen atom(s), nitrogen atom(s),and/or sulfur atom(s) in the parent compound, however.

The phrase “substituted aryl group” has the same meaning with respect tounsubstituted aryl groups that substituted alkyl groups had with respectto unsubstituted alkyl groups. However, a substituted aryl group alsoincludes aryl groups in which one of the aromatic carbons is bonded toone of the non-carbon or non-hydrogen atoms described above and alsoincludes aryl groups in which one or more aromatic carbons of the arylgroup is bonded to a substituted and/or unsubstituted alkyl, alkenyl, oralkynyl group as defined herein. This includes bonding arrangements inwhich two carbon atoms of an aryl group are bonded to two atoms of analkyl, alkenyl, or alkynyl group to define a fused ring system (e.g.dihydronaphthyl or tetrahydronaphthyl). Thus, the phrase “substitutedaryl” includes, but is not limited to tolyl, and hydroxyphenyl amongothers.

The phrase “unsubstituted alkenyl” refers to straight and branched chainand cyclic groups such as those described with respect to unsubstitutedalkyl groups as defined above, except that at least one double bondexists between two carbon atoms. Examples include, but are not limitedto vinyl, —CH═C(H)(CH₃), —CH═C(CH₃)₂, —C(CH₃)═C(H)₂, —C(CH₃)═C(H)(CH₃),—C(CH₂CH₃)═CH₂, cyclohexenyl, cyclopentenyl, cyclohexadienyl,butadienyl, pentadienyl, and hexadienyl among others.

The phrase “substituted alkenyl” has the same meaning with respect tounsubstituted alkenyl groups that substituted alkyl groups had withrespect to unsubstituted alkyl groups. A substituted alkenyl groupincludes alkenyl groups in which a non-carbon or non-hydrogen atom isbonded to a carbon double bonded to another carbon and those in whichone of the non-carbon or non-hydrogen atoms is bonded to a carbon notinvolved in a double bond to another carbon.

The phrase “unsubstituted alkynyl” refers to straight and branched chaingroups such as those described with respect to unsubstituted alkylgroups as defined above, except that at least one triple bond existsbetween two carbon atoms. Examples include, but are not limited to—C≡C(H), —C≡C(CH₃), —C≡C(CH₂CH₃), —C(H₂)C≡C(H), —C(H)₂C≡C(CH₃), and—C(H)₂C≡C(CH₂CH₃) among others.

The phrase “substituted alkynyl” has the same meaning with respect tounsubstituted alkynyl groups that substituted alkyl groups had withrespect to unsubstituted alkyl groups. A substituted alkynyl groupincludes alkynyl groups in which a non-carbon or non-hydrogen atom isbonded to a carbon triple bonded to another carbon and those in which anon-carbon or non-hydrogen atom is bonded to a carbon not involved in atriple bond to another carbon.

The phrase “unsubstituted aralkyl” refers to unsubstituted alkyl groupsas defined above in which a hydrogen or carbon bond of the unsubstitutedalkyl group is replaced with a bond to an aryl group as defined above.For example, methyl (—CH₃) is an unsubstituted alkyl group. If ahydrogen atom of the methyl group is replaced by a bond to a phenylgroup, such as if the carbon of the methyl were bonded to a carbon ofbenzene, then the compound is an unsubstituted aralkyl group (i.e., abenzyl group). Thus the phrase includes, but is not limited to, groupssuch as benzyl, diphenylmethyl, and 1-phenylethyl (—CH(C₆H₅)(CH₃)) amongothers.

The phrase “substituted aralkyl” has the same meaning with respect tounsubstituted aralkyl groups that substituted aryl groups had withrespect to unsubstituted aryl groups. However, a substituted aralkylgroup also includes groups in which a carbon or hydrogen bond of thealkyl part of the group is replaced by a bond to a non-carbon or anon-hydrogen atom. Examples of substituted aralkyl groups include, butare not limited to, —CH₂C(═O)(C₆H₅), and —CH₂(2-methylphenyl) amongothers.

The phrase “unsubstituted heterocyclyl” refers to both aromatic andnonaromatic ring compounds including monocyclic, bicyclic, andpolycyclic ring compounds such as, but not limited to, quinuclidyl,containing 3 or more ring members of which one or more is a heteroatomsuch as, but not limited to, N, O, and S. Although the phrase“unsubstituted heterocyclyl” includes condensed heterocyclic rings suchas benzimidazolyl, it does not include heterocyclyl groups that haveother groups such as alkyl or halo groups bonded to one of the ringmembers as compounds such as 2-methylbenzimidazolyl are substitutedheterocyclyl groups. Examples of heterocyclyl groups include, but arenot limited to: unsaturated 3 to 8 membered rings containing 1 to 4nitrogen atoms such as, but not limited to pyrrolyl, pyrrolinyl,imidazolyl, pyrazolyl, pyridinyl, dihydropyridinyl, pyrimidyl,pyrazinyl, pyridazinyl, triazolyl (e.g. 4H-1,2,4-triazolyl,1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl etc.), tetrazolyl, (e.g.1H-tetrazolyl, 2H tetrazolyl, etc.); saturated 3 to 8 membered ringscontaining 1 to 4 nitrogen atoms such as, but not limited to,pyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl; condensedunsaturated heterocyclic groups containing 1 to 4 nitrogen atoms suchas, but not limited to, indolyl, isoindolyl, indolinyl, indolizinyl,benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl;unsaturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1to 3 nitrogen atoms such as, but not limited to, oxazolyl, isoxazolyl,oxadiazolyl (e.g. 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl,1,2,5-oxadiazolyl, etc.); saturated 3 to 8 membered rings containing 1to 2 oxygen atoms and 1 to 3 nitrogen atoms such as, but not limited to,morpholinyl; unsaturated condensed heterocyclic groups containing 1 to 2oxygen atoms and 1 to 3 nitrogen atoms, for example, benzoxazolyl,benzoxadiazolyl, benzoxazinyl (e.g. 2H-1,4-benzoxazinyl etc.);unsaturated 3 to 8 membered rings containing 1 to 3 sulfur atoms and 1to 3 nitrogen atoms such as, but not limited to, thiazolyl,isothiazolyl, thiadiazolyl (e.g. 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, etc.); saturated 3 to 8 memberedrings containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms such as,but not limited to, thiazolodinyl; saturated and unsaturated 3 to 8membered rings containing 1 to 2 sulfur atoms such as, but not limitedto, thienyl, dihydrodithionyl, dihydrodithionyl, tetrahydrothiophene,tetrahydrothiopyran; unsaturated condensed heterocyclic rings containing1 to 2 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limitedto, benzothiazolyl, benzothiadiazolyl, benzothiazinyl (e.g.2H-1,4-benzothiazinyl, etc.), dihydrobenzothiazinyl (e.g.2H-3,4-dihydrobenzothiazinyl, etc.), unsaturated 3 to 8 membered ringscontaining oxygen atoms such as, but not limited to furyl; unsaturatedcondensed heterocyclic rings containing 1 to 2 oxygen atoms such asbenzodioxolyl (e.g. 1,3-benzodioxoyl, etc.); unsaturated 3 to 8 memberedrings containing an oxygen atom and 1 to 2 sulfur atoms such as, but notlimited to, dihydrooxathiinyl; saturated 3 to 8 membered ringscontaining 1 to 2 oxygen atoms and 1 to 2 sulfur atoms such as1,4-oxathiane; unsaturated condensed rings containing 1 to 2 sulfuratoms such as benzothienyl, benzodithiinyl; and unsaturated condensedheterocyclic rings containing an oxygen atom and 1 to 2 oxygen atomssuch as benzoxathiinyl. Heterocyclyl group also include those describedabove in which one or more S atoms in the ring is double-bonded to oneor two oxygen atoms (sulfoxides and sulfones). For example, heterocyclylgroups include tetrahydrothiophene oxide and tetrahydrothiophene1,1-dioxide. Preferred heterocyclyl groups contain 5 or 6 ring members.More preferred heterocyclyl groups include morpholine, piperazine,piperidine, pyrrolidine, imidazole, pyrazole, 1,2,3-triazole,1,2,4-triazole, tetrazole, thiophene, thiomorpholine, thiomorpholine inwhich the S atom of the thiomorpholine is bonded to one or more O atoms,pyrrole, homopiperazine, oxazolidin-2-one, pyrrolidin-2-one, oxazole,quinuclidine, thiazole, isoxazole, furan, and tetrahydrofuran.

The phrase “substituted heterocyclyl” refers to an unsubstitutedheterocyclyl group as defined above in which one or more of the ringmembers is bonded to a non-hydrogen atom such as described above withrespect to substituted alkyl groups and substituted aryl groups.Examples include, but are not limited to, 2-methylbenzimidazolyl,5-methylbenzimidazolyl, 5-chlorobenzthiazolyl, 1-methyl piperazinyl,2-phenoxy-thiophene, and 2-chloropyridinyl among others. In addition,substituted heterocyclyl groups also include heterocyclyl groups inwhich the bond to the non-hydrogen atom is a bond to a carbon atom thatis part of a substituted and unsubstituted aryl, substituted andunsubstituted arylalkyl, or unsubstituted heterocyclyl group. Examplesinclude but are not limited to 1-benzylpiperdinyl,3-phenythiomorpholinyl, 3-(pyrrolidin-1-yl)-pyrrolidinyl, and4-(piperidin-1-yl)-piperidinyl.

The phrase “unsubstituted heterocyclylalkyl” refers to unsubstitutedalkyl groups as defined above in which a hydrogen or carbon bond of theunsubstituted alkyl group is replaced with a bond to a heterocyclylgroup as defined above. For example, methyl (—CH₃) is an unsubstitutedalkyl group. If a hydrogen atom of the methyl group is replaced by abond to a heterocyclyl group, such as if the carbon of the methyl werebonded to carbon 2 of pyridine (one of the carbons bonded to the N ofthe pyridine) or carbons 3 or 4 of the pyridine, then the compound is anunsubstituted heterocyclylalkyl group.

The phrase “substituted heterocyclylalkyl” has the same meaning withrespect to unsubstituted heterocyclylalkyl groups that substitutedaralkyl groups had with respect to unsubstituted aralkyl groups.However, a substituted heterocyclylalkyl group also includes groups inwhich a non-hydrogen atom is bonded to a heteroatom in the heterocyclylgroup of the heterocyclylalkyl group such as, but not limited to, anitrogen atom in the piperidine ring of a piperidinylalkyl group. Inaddition, a substituted heterocyclylalkyl group also includes groups inwhich a carbon bond or a hydrogen bond of the alkyl part of the group isreplaced by a bond to a substituted and unsubstituted aryl orsubstituted and unsubstituted arylalkyl group. Examples include but arenot limited to phenyl-(piperidin-1-yl)-methyl andphenyl-(morpholin-4-yl)-methyl.

The phrase “unsubstituted alkoxy” refers to a hydroxyl group (—OH) inwhich the bond to the hydrogen atom is replaced by a bond to a carbonatom of an otherwise unsubstituted alkyl group as defined above.

The phrase “substituted alkoxy” refers to a hydroxyl group (—OH) inwhich the bond to the hydrogen atom is replaced by a bond to a carbonatom of an otherwise substituted alkyl group as defined above.

The term “protected” with respect to hydroxyl groups, amine groups, andsulfhydryl groups refers to forms of these functionalities which areprotected from undesirable reaction with a protecting group known tothose skilled in the art such as those set forth in Protective Groups inOrganic Synthesis, Greene, T. W.; Wuts, P. G. M., John Wiley & Sons, NewYork, N.Y., (3rd Edition, 1999) which can be added or removed using theprocedures set forth therein. Examples of protected hydroxyl groupsinclude, but are not limited to, silyl ethers such as those obtained byreaction of a hydroxyl group with a reagent such as, but not limited to,t-butyldimethyl-chlorosilane, trimethylchlorosilane,triisopropylchlorosilane, triethylchlorosilane; substituted methyl andethyl ethers such as, but not limited to methoxymethyl ether,methylthiomethyl ether, benzyloxymethyl ether, t-butoxymethyl ether,2-methoxyethoxymethyl ether, tetrahydropyranyl ethers, 1-ethoxyethylether, allyl ether, benzyl ether; esters such as, but not limited to,benzoylformate, formate, acetate, trichloroacetate, and trifluoracetate.Examples of protected amine groups include, but are not limited to,amides such as, formamide, acetamide, trifluoroacetamide, and benzamide;imides, such as phthalimide, and dithiosuccinimide; and others. Examplesof protected sulfhydryl groups include, but are not limited to,thioethers such as S-benzyl thioether, and S-4-picolyl thioether;substituted S-methyl derivatives such as hemithio, dithio and aminothioacetals; and others.

A “pharmaceutically acceptable salt” includes a salt with an inorganicbase, organic base, inorganic acid, organic acid, or basic or acidicamino acid. As salts of inorganic bases, the invention includes, forexample, alkali metals such as sodium or potassium; alkaline earthmetals such as calcium and magnesium or aluminum; and ammonia. As saltsof organic bases, the invention includes, for example, trimethylamine,triethylamine, pyridine, picoline, ethanolamine, diethanolamine, andtriethanolamine. As salts of inorganic acids, the instant inventionincludes, for example, hydrochloric acid, hydroboric acid, nitric acid,sulfuric acid, and phosphoric acid. As salts of organic acids, theinstant invention includes, for example, formic acid, acetic acid,trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, maleicacid, citric acid, succinic acid, malic acid, methanesulfonic acid,ethanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid.As salts of basic amino acids, the instant invention includes, forexample, arginine, lysine and ornithine. Acidic amino acids include, forexample, aspartic acid and glutamic acid.

Prodrugs, as used in the context of the instant invention, includesthose derivatives of the instant compounds which undergo in vivometabolic biotransformation, by enzymatic or nonenzymatic processes,such as hydrolysis, to form a compound of the invention. Typicalprodrugs include ester and ether moieties. Prodrugs can be employed toimprove pharmaceutical or biological properties, as for examplesolubility, melting point, stability and related physicochemicalproperties, absorption, pharmacodynamics and other delivery-relatedproperties.

In accordance with I.U.P.A.C. nomenclature, the carbon atoms of thefused four-ring system of the present invention are numbered as follows:

Tautomers refers to isomeric forms of a compound that are in equilibriumwith each other. The concentrations of the isomeric forms will depend onthe environment the compound is found in and may be different dependingupon, for example, whether the compound is a solid or is in an organicor aqueous solution. For example, in aqueous solution, ketones aretypically in equilibrium with their enol forms. Thus, ketones and theirenols are referred to as tautomers of each other. As readily understoodby one skilled in the art, a wide variety of functional groups and otherstructures may exhibit tautomerism, and all tautomers of compoundshaving structures I, II, or III are within the scope of the presentinvention.

Compounds of the present invention include enriched or resolved opticalisomers at any or all asymmetric atoms as are apparent from thedepictions. Both racemic and diastereomeric mixtures, as well as theindividual optical isomers can be isolated or synthesized so as to besubstantially free of their enantiomeric or diastereomeric partners, andthese are all within the scope of the invention.

In accordance with one aspect of the invention, there are providedcompounds of structure I, prodrugs thereof, pharmaceutically acceptablesalts thereof, stereoisomers thereof, tautomers thereof, and solvatesthereof.

In compounds of structure I, A is —C(O)—, ═CR⁹—, or —CR⁹R¹⁰—; E is—C(O)—, ═CR⁵—, or —CR⁵R⁶—, wherein A and E are not both —C(O)—; G is—C(O)—, ═CR³—, or —CR³R⁴ and K is —C(O)—, ═CR¹—, or —CR¹R²—.

In certain embodiments of compounds having the structure I, A is—CR⁹R¹⁰—, E is —CR⁵R⁶—, G is —CR³R⁴—, or K is —C(OR¹¹)R²—. In other suchembodiments, A is —CR⁹R¹⁰— and K is —CR¹R²—. In further embodiments, Eis —CR⁵R⁶—, G is —CR³R⁴—, and K is —CR¹R²—. In still furtherembodiments, A is —CR⁹R¹⁰—, E is —CR⁵R⁶—, G is —CR³R⁴—, and K is—CR¹R²—. In other embodiments, A is —C(O)—, E is —C(O)—, G is —C(O)—, orK is —C(O)—.

The present invention also contemplates compounds of structure I havingdouble bonds at particular positions as set forth below. Thus, in someembodiments, A is ═CR⁹—, G is ═CR³—, or E is ═CR⁵—. In other embodimentsA is ═CR⁹— and E is ═CR⁵—.

In compounds of structure I, R¹ is selected from the group consisting of—OR¹¹, substituted and unsubstituted alkyl, substituted andunsubstituted alkene, substituted and unsubstituted alkyne, substitutedand unsubstituted aryl, substituted and unsubstituted arylalkyl,substituted and unsubstituted heterocyclyl, substituted andunsubstituted heterocyclylalkyl, —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹²,—C(S)—R¹², —C(S)—OR¹², —NR¹²R¹³, —NR¹²—C(O)—R¹³, —NR¹²—C(O)—OR¹³,—NR¹²—C(O)—NR¹²R¹³, and —S(O)₀₋₂—R¹². In some embodiments, R¹ isselected from the group consisting of —OR¹¹, substituted andunsubstituted alkyl, —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹²,—C(S)—OR¹², —NR¹²R¹³, —NR¹²—C(O)—R¹³, —NR¹²—C(O)—OR¹³,—NR¹²—C(O)—NR¹²R¹³, and —S(O)₀₋₂—R¹². In other embodiments, R¹ isselected from the group consisting of —OR¹¹, substituted andunsubstituted alkyl, —NR¹²R¹³, —NR¹²—C(O)—R¹³, and —NR¹²—C(O)—OR¹³. Inyet other embodiments, R¹ is —OR¹¹.

In compounds of structure I, R² is selected from the group consisting of—H, substituted and unsubstituted lower alkyl, substituted andunsubstituted lower alkene, and substituted and unsubstituted loweralkyne. In other embodiments, R² is —H.

In compounds of structure I, R³ and R⁵ are independently selected fromthe group consisting of —H, —F, —Cl, —Br, —I, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,substituted and unsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵,—NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted loweralkoxy, and —S(O)₀₋₂R¹⁴. In some embodiments, R³ and R⁵ areindependently selected from the group consisting of —H, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkyne,—CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH, andsubstituted and unsubstituted lower alkoxy. In other embodiments, R³ andR⁵ are independently selected from the group consisting of —H,substituted and unsubstituted lower alkyl, substituted and unsubstitutedlower alkyne, —OH, and substituted and unsubstituted lower alkoxy. Instill other embodiments, R³ or R⁵ is —H, or both are —H.

In compounds of structure I, R⁴ and R⁶ are independently selected fromthe group consisting of —H, —F, —Cl, —Br, —I, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,substituted and unsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵,—NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted loweralkoxy, and —S(O)₀₋₂R¹⁴.

In compounds of structure I, R⁷ and R⁸ are independently selected fromthe group consisting of —H and substituted and unsubstituted loweralkyl. In some such embodiments, R⁷ and R⁸ are independently selectedfrom unsubstituted lower alkyl. In other embodiments, R⁷ or R⁸ ismethyl, or both are methyl.

In compounds of structure I, R⁹ and R¹⁰ are independently selected fromthe group consisting of substituted and unsubstituted lower alkyl,substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —OH, wherein R⁹ and R¹⁰ are not both —OH,substituted and unsubstituted lower alkoxy, and substituted andunsubstituted —S(O)₀₋₂(lower alkyl), or R⁹ and R¹⁰, together with thecarbon to which they are attached, form a 5-, 6-, or 7-memberheterocyclyl or cycloalkyl group. In some embodiments R⁹ is a —OH orsubstituted alkoxy, such as an acetyl and the like.

In other embodiments of compounds having structure I, R⁹ and R¹⁰,together with the carbon to which they are attached, form a 5-, 6-, or7-member heterocyclyl or cycloalkyl group. Certain such embodiments,have the structure

wherein,

X and Y are independently selected from the group consisting of —NR¹⁴—,—O—, —S—, and substituted and unsubstituted C₁ alkyl;

Z is substituted or unsubstituted C₂₋₄ alkyl or substituted orunsubstituted —(CR¹⁴R¹⁵)₂₋₃—; and

R² is as defined herein.

In other embodiments, R⁹ and R¹⁰, together with the carbon to which theyare attached, form a 5-, 6-, or 7-member heterocyclyl. Typically theheterocycle is a 5- or 6-member heterocycle such as a ketal or thioketal

In compounds of structure I, R¹¹ is selected from the group consistingof —H, substituted and unsubstituted alkyl, substituted andunsubstituted alkene, substituted and unsubstituted alkyne, substitutedand unsubstituted aryl, substituted and unsubstituted arylalkyl,substituted and unsubstituted heterocyclyl, substituted andunsubstituted heterocyclylalkyl, —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹²,—C(S)—R¹², —NR¹²R¹³, —S(O)₂—R¹², —S(O)₂—OR¹², and —P(O)(OR¹²)(OR¹³)₀₋₁.In some embodiments, R¹¹ is selected from the group consisting ofsubstituted and unsubstituted alkyl, substituted and unsubstitutedalkene, substituted and unsubstituted alkyne, substituted andunsubstituted aryl, substituted and unsubstituted arylalkyl, substitutedand unsubstituted heterocyclyl, substituted and unsubstitutedheterocyclylalkyl, —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹²,—NR¹²R¹³, —S(O)₂—R¹², —S(O)₂—OR¹², or —P(O)(OR¹²)(OR¹³)₀₋₁. In otherembodiments, R¹¹ is selected from the group consisting of —H,substituted and unsubstituted alkyl, —C(O)—R¹², —C(O)—NR¹²R¹³, and—C(O)—OR¹². In still other embodiments, R¹¹ is selected from the groupconsisting of —H, —C(O)—R¹² and —C(O)—OR¹². In some embodiments, R¹¹ is—C(O)—R¹².

In compounds of structure I, R¹² and R¹³ are, at each occurrence,independently selected from the group consisting of —H, substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, and substituted and unsubstituted heterocyclylalkyl. Insome embodiments, R¹² and R¹³ are, at each occurrence, independentlyselected from the group consisting of —H, substituted and unsubstitutedalkyl, substituted and unsubstituted alkene, and substituted andunsubstituted alkyne. In some such embodiments, R¹² is selected from thegroup consisting of —H and substituted and unsubstituted lower alkyl. Instill other embodiments where R¹¹ is —C(O)—R¹², R¹² is unsubstitutedlower alkyl. In some embodiments, R¹³ is —H.

In compounds of structure I, R¹⁴ and R¹⁵ are, at each occurrence,independently selected from the group consisting of substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,substituted and unsubstituted lower alkyne, substituted andunsubstituted C₆₋₁₀ aryl, and substituted and unsubstituted C₇₋₁₂arylalkyl.

In compounds of structure I, R and R′ are, at each occurrence,independently selected from the group consisting of —F, —Cl, —Br, —I,substituted and unsubstituted lower alkyl, substituted and unsubstitutedlower alkene, substituted and unsubstituted lower alkyne, —CN, —COOR¹⁴,—C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH, substituted andunsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴. In other embodiments R andR′ are, at each occurrence, independently selected from the groupconsisting of —F, —Cl, —Br, —I, substituted and unsubstituted loweralkyl, substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —OH, and substituted and unsubstituted loweralkoxy. In still other embodiments, R and R′ are, at each occurrence,independently selected from the group consisting of —F and substitutedand unsubstituted lower alkyl, —OH, and substituted and unsubstitutedlower alkoxy.

In compounds of structure I, typically, n and n′ are independently 0, 1,or 2. In some embodiments, n and n′ are independently 1 or 2. In otherembodiments, n and n′ are independently 0 or 1. Thus, the A ring of thefused four-ring system of structure I may have one or two R substituentsat position 1, one or two R substituents at position 2, and so forth.Alternatively, the A ring may have two R substituents at differentpositions such as position 2 and 4. Similarly, in some embodiments theB-ring may have one or two R′ substituents at position 6 or a single R′at position 9, among other embodiments.

In compounds of structure I, the dashed lines in structure I representcarbon-carbon double bonds or carbon-carbon single bonds containedwithin the fused four-ring system, such that the compound comprises a1,6-diene, 1,7-diene, 1,8-diene, 1,15-diene, 1,16-diene, 4,8-diene,3,16-diene, 1,3,5-triene, 1,3,16-triene, 1,5,7-triene, 1,5,15-triene,1,8,15-triene, 1,5,16-triene, or 1,5,7,15-tetraene, within the fusedfour-ring system. The 1,8-diene includes both 1,8(9)- and1,8(14)-dienes, while the 1,8,15-triene includes 1,8(9)- and1,8(14)-trienes. In some embodiments, the compound comprises a1,6-diene, 1,7-diene, 1,8-diene, 1,15-diene, 1,16-diene, 4,8-diene, or3,16-diene, within the fused four-ring system. In other embodiments, thecompound comprises a 1,3,5-triene, 1,3,16-triene, 1,5,7-triene,1,5,15-triene, 1,8,15-triene, 1,5,16-triene, or 1,5,7,15-tetraene withinthe fused four-ring system.

In another aspect of the invention, there are provided compounds ofstructure II, prodrugs thereof, pharmaceutically acceptable saltsthereof, stereoisomers thereof, tautomers thereof, or solvates thereof.

In compounds of structure II, A is —C(O)— or —CR⁹R¹⁰—; E is —C(O)— or—CR⁵R⁶—, wherein A and E are not both —C(O)—; G is —C(O)—, ═CR³—, or—CR³R⁴—; and K is ═C(OR¹¹)—, or —C(OR¹¹)R²—.

In certain embodiments of compounds having the structure II, A is—CR⁹R¹⁰—, E is —CR⁵R⁶—, G is —CR³R⁴—, or K is —C(OR¹¹)R²—. In other suchembodiments, A is —CR⁹R¹⁰—, and K is —C(OR¹¹)R¹—. In furtherembodiments, E is —CR⁵R⁶—, G is —CR³R⁴—, and K is —C(OR¹¹)R²—. In stillfurther embodiments, A is —CR⁹R¹⁰—, E is —CR⁵R⁶—, G is —CR³R⁴—, and K is—C(OR¹¹)R²—. In other embodiments, A is —C(O)—, E is —C(O)—, or G is—C(O)—. Alternatively, G is ═CR³—.

In compounds of structure II, R² is selected from the group consistingof —H, substituted and unsubstituted lower alkyl, substituted andunsubstituted lower alkene, and substituted and unsubstituted loweralkyne. In other embodiments, R² is —H.

In compounds of structure II, R³ and R⁵ are independently selected fromthe group consisting of —H, —F, —Cl, —Br, —I, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,substituted and unsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵,—NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted loweralkoxy, and —S(O)₀₋₂R¹⁴. In some embodiments, R³ and R⁵ areindependently selected from the group consisting of —H, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkyne,—CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH, andsubstituted and unsubstituted lower alkoxy. In other embodiments, R³ andR⁵ are independently selected from the group consisting of —H,substituted and unsubstituted lower alkyl, substituted and unsubstitutedlower alkyne, —OH, and substituted and unsubstituted lower alkoxy. Instill other embodiments, R³ or R⁵ is —H, or both are —H.

In compounds of structure II, R⁴ and R⁶ are independently selected fromthe group consisting of —H, —F, —Cl, —Br, —I, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,substituted and unsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵,—NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted loweralkoxy, and —S(O)₀₋₂R¹⁴.

In compounds of structure II, R⁷ and R⁸ are independently selected fromthe group consisting of —H and substituted and unsubstituted loweralkyl. In some such embodiments, R⁷ and R⁸ are independently selectedfrom unsubstituted lower alkyl. In other embodiments, R⁷ or R⁸ ismethyl, or both are methyl.

In compounds of structure II, R⁹ and R¹⁰ are independently selected fromthe group consisting of substituted and unsubstituted lower alkyl,substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —OH, wherein R⁹ and R¹⁰ are not both —OH,substituted and unsubstituted lower alkoxy, and substituted andunsubstituted —S(O)₀₋₂(lower alkyl); or R⁹ and R¹⁰, together with thecarbon to which they are attached, form a 5-, 6-, or 7-memberheterocyclyl or cycloalkyl group. In some such embodiments, R⁹ and R¹⁰,together with the carbon to which they are attached, form a 5-, 6-, or7-member heterocyclyl or cycloalkyl group. In some embodiments, R⁹ andR¹⁰, together with the carbon to which they are attached, form a 5-, 6-,or 7-member heterocyclyl. Typically the heterocycle is a 5- or 6-memberheterocycle such as a ketal.

In compounds of structure II, R¹¹ is selected from the group consistingof substituted and unsubstituted alkyl, substituted and unsubstitutedalkene, substituted and unsubstituted alkyne, substituted andunsubstituted aryl, substituted and unsubstituted arylalkyl, substitutedand unsubstituted heterocyclyl, substituted and unsubstitutedheterocyclylalkyl, —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹²,—NR¹²R¹³, —S(O)₂—R¹², —S(O)₂—OR¹², and —P(O)(OR¹²)(OR¹³)₀₋₁. In someembodiments, R¹¹ is selected from the group consisting of substitutedand unsubstituted alkyl, —C(O)—R¹², —C(O)—NR¹²R¹³, and —C(O)—OR¹². Inother embodiments, R¹¹ is selected from the group consisting of—C(O)—R¹² and —C(O)—OR¹². In still other embodiments, R¹¹ is —C(O)—R¹².

In compounds of structure II, R¹² and R¹³ are, at each occurrence,independently selected from the group consisting of —H, substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, and substituted and unsubstituted heterocyclylalkyl. Insome embodiments, R¹² and R¹³ are, at each occurrence, independentlyselected from the group consisting of —H, substituted and unsubstitutedalkyl, substituted and unsubstituted alkene, and substituted andunsubstituted alkyne. In some such embodiments, R¹² is selected from thegroup consisting of —H and substituted and unsubstituted lower alkyl. Instill other embodiments where R¹¹ is —C(O)—R¹², R¹² is unsubstitutedlower alkyl. In some embodiments, R¹³ is —H.

In compounds of structure II, R¹⁴ and R¹⁵ are, at each occurrence,independently selected from the group consisting of substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,substituted and unsubstituted lower alkyne, substituted andunsubstituted C₆₋₁₀ aryl, and substituted and unsubstituted C₇₋₁₂arylalkyl.

In compounds of structure II, R and R′ are, at each occurrence,independently selected from the group consisting of —F, —Cl, —Br, —I,substituted and unsubstituted lower alkyl, substituted and unsubstitutedlower alkene, substituted and unsubstituted lower alkyne, —CN, —COOR¹⁴,—C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH, substituted andunsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴. In other embodiments R andR′ are, at each occurrence, independently selected from the groupconsisting of —F, —Cl, —Br, —I, substituted and unsubstituted loweralkyl, substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —OH, and substituted and unsubstituted loweralkoxy. In still other embodiments, R and R′ are, at each occurrence,independently selected from the group consisting of —F and substitutedand unsubstituted lower alkyl, —OH, and substituted and unsubstitutedlower alkoxy.

In compounds of structure II, typically, n and n′ are independently 0,1, or 2. In some embodiments, n and n′ are independently 1 or 2. Inother embodiments, n and n′ are independently 0 or 1. Thus, the A ringof the fused four-ring system of structure I may have one or two Rsubstituents at position 1, one or two R substituents at position 2, andso forth. Alternatively, the A ring may have two R substituents atdifferent positions such as position 2 and 4. Similarly, in someembodiments the B-ring may have one or two R′ substituents at position 6or a single R′ at position 9, among other embodiments.

In compounds of structure II, the dashed lines in structure II representcarbon-carbon double bonds or carbon-carbon single bonds containedwithin the fused four-ring system, such that the compound comprises a1,3-diene, 1,5-diene, or 1,4,6-triene within the fused four-ring system.In some embodiments, the compound comprises a 1,3-diene or 1,4,6-trienewithin the fused four-ring system. In other embodiments, the compoundcomprises a 1,5-diene within the fused four-ring system. Certain suchembodiments have the structure IIA:

wherein,

X and Y are independently selected from the group consisting of —NR¹⁴—,—O—, —S—, and substituted and unsubstituted C₁ alkyl;

Z is substituted or unsubstituted C₂₋₄ alkyl, e.g., —CH₂—CH₂— or—CH₂—CH₂—CH₂—, or substituted or unsubstituted —(CR¹⁴R¹⁵)₂₋₃—; and

R² is as previously defined.

Thus, the invention provides compounds having the structures:

In still another aspect of the invention, there are provided methods oftreating or preventing a condition mediated by an androgen receptorcomprising administering to a subject in need thereof, an effectiveamount of a compound having the structure III, prodrugs thereof,pharmaceutically acceptable salts thereof, stereoisomers thereof,tautomers thereof, or solvates thereof.

In compounds of structure III

A is —C(O)—, ═CR⁹—, or —CR⁹R¹⁰—;

E is —C(O)—, ═CR⁵—, or —CR⁵R⁶—, wherein A and E are not both —C(O);

G is —C(O)—, ═CR³—, or —CR³R⁴—;

K is —C(O)—, ═CR¹—, or —CR¹R²—;

R¹ is selected from the group consisting of —OR¹¹, substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, substituted and unsubstituted heterocyclylalkyl,—C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹², —C(S)—OR¹², —NR¹²R¹³,—NR¹²—C(O)—R¹³, —NR¹²—C(O)—OR¹³, —NR¹²—C(O)—NR¹²R¹³, and —S(O)₀₋₂—R¹²;

R² is selected from the group consisting of —H, substituted andunsubstituted lower alkyl, substituted and unsubstituted lower alkene,and substituted and unsubstituted lower alkyne;

R³ and R⁵ are independently selected from the group consisting of —H,—F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl, substitutedand unsubstituted lower alkene, substituted and unsubstituted loweralkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH,substituted and unsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴;

R⁴ and R⁶ are independently selected from the group consisting of —H,—F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl, substitutedand unsubstituted lower alkene, substituted and unsubstituted loweralkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵, —NR¹⁴—C(O)—R¹⁵, —OH,substituted and unsubstituted lower alkoxy, and —S(O)₀₋₂R¹⁴;

R⁷ and R³ are independently selected from the group consisting of —H andsubstituted and unsubstituted lower alkyl group;

R⁹ and R¹⁰ are independently selected from the group consisting ofsubstituted and unsubstituted lower alkyl, substituted and unsubstitutedlower alkene, substituted and unsubstituted lower alkyne, —OH, whereinR⁹ and R¹⁰ are not both —OH, substituted and unsubstituted lower alkoxy,and substituted and unsubstituted —S(O)₀₋₂(lower alkyl), or R⁹ and R¹⁰,together with the carbon to which they are attached, form a 5-, 6-, or7-member heterocyclyl or cycloalkyl group;

R¹¹ is selected from the group consisting of —H, substituted andunsubstituted alkyl, substituted and unsubstituted alkene, substitutedand unsubstituted alkyne, substituted and unsubstituted aryl,substituted and unsubstituted arylalkyl, substituted and unsubstitutedheterocyclyl, substituted and unsubstituted heterocyclylalkyl,—C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹², —C(S)—R¹², —NR¹²R¹³, S(O)₂—R¹²,—S(O)₂—OR¹², or —P(O)(OR¹²)(OR¹³)₀₋₁;

R¹² and R¹³ are, at each occurrence, independently selected from thegroup consisting of —H, substituted and unsubstituted alkyl, substitutedand unsubstituted alkene, substituted and unsubstituted alkyne,substituted and unsubstituted aryl, substituted and unsubstitutedarylalkyl, substituted and unsubstituted heterocyclyl, and substitutedand unsubstituted heterocyclylalkyl;

R¹⁴ and R¹⁵ are, at each occurrence, independently selected from thegroup consisting of substituted and unsubstituted lower alkyl,substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, substituted and unsubstituted C₆₋₁₀ aryl,and substituted and unsubstituted C₇₋₁₂ arylalkyl;

R and R′ are, at each occurrence, independently selected from the groupconsisting of —F, —Cl, —Br, —I, substituted and unsubstituted loweralkyl, substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted lower alkoxy, and—S(O)₀₋₂R¹⁴;

n and n′ are independently 0, 1, or 2; and

wherein the dashed lines in structure III represent carbon-carbon doublebonds or carbon-carbon single bonds contained within the fused four-ringsystem, such that the compound comprises a 1,3-diene, 1,5-diene,1,6-diene, 1,7-diene, 1,8-diene, 1,15-diene, 1,16-diene, 3,16-diene,4,8-diene, 1,3,5-triene, 1,4,6-triene, 1,3,16-triene, 1,5,7-triene,1,5,15-triene, 1,8,15-triene, 1,5,16-triene, or 1,5,7,15-tetraene,within the fused four-ring system.

In the compounds described herein, the hydrogen atoms at the 8, 9 and 14positions respectively are typically in the β-, α- and α-configurations.

In some embodiments of methods of treating or preventing a conditionmediated by an androgen receptor, the condition is prostate cancer, andin particular, prostate cancer at an androgen-independent stage. Inother embodiments, the condition is antiandrogen induced withdrawalsyndrome, and the subject may be afflicted with prostate cancer. Instill further embodiments, the condition is benign prostatichypertrophy, hirsutism, acne, androgenic alopecia, or ovulatorydysfunction in hyperandrogenic women, such as, for example, polycysticovary syndrome patients. In some embodiments, the compounds disclosedherein are used to ameliorate and/or slow the progression of one or moreof these conditions.

In other embodiments of methods of treating, preventing or slowing theprogression of a condition mediated by an androgen receptor, thecompound comprises a 1,5-diene within the fused four-ring system. Inother such embodiments, K is —CR¹R²—, R¹ is —OR¹¹, or R¹¹ is —H,substituted or unsubstituted alkyl, —C(O)—R¹², —C(O)—NR¹²R¹³, or—C(O)—OR¹². In further embodiments, the compound comprises a 1,5-dienewithin the fused four-ring system, and R¹¹ is —H, or —C(O)—R¹². Thus,the invention provides methods of treating, preventing or ameliorating acondition mediated by an androgen receptor using compounds disclosedherein. Such compounds include ADEK,3β-hydroxyandrosta-1,5-dien-17,17-ethylene ketal,3β-hydroxyandrosta-1,5-dien-17-one and3β-acetoxyandrosta-1,5-dien-17-one.

While not wishing to be bound by any theory, it is believed that thecompounds of the present invention are advantageously used in treatingandrogen-receptor mediated conditions because, among other things, theyinhibit the activity of Adiol. The latter compound is unique amongnaturally occurring androgens in that its transactivation of theandrogen receptor is not inhibited by previously known antiandrogenssuch as hydroxyflutamide or bicalutamide. Thus, Adiol activity cancontribute to, e.g., androgen-independent prostate cancer. In contrast,as shown in Example 6, inventive compounds repress Adiol-induced ARtranscription and would be expected to show efficacy againstandrogen-independent prostate cancer.

The present invention thus provides methods of inhibiting androgenreceptors in vitro or in vivo comprising contacting an androgen receptorwith an effective amount of a compound, e.g., a compound having thestructure I, II or III. In some embodiments of such methods, thetransactivation of androgen receptor is suppressed. In otherembodiments, the androgen receptor is mutant or native androgenreceptor. Such embodiments include methods to modulate the biologicalactivity and/or the level of androgen receptor activity, e.g., in humansor mammals who have, or who are disposed to develop, an androgenreceptor related condition or symptom. Such modulation can be effectedin cells in vitro or in vivo. Compounds such as those described herein,or other androgen receptor modulators, e.g., as described in U.S. Pat.No. 6,645,974 B2, 6,569,896 B2, 6,696,459 B1 or 6,710,037 B2, can becharacterized by their capacity to antagonize the activity of androgenreceptor agonists such as adiol. The capacity of any selected testcompound to modulate or antagonize androgen receptor activity or levelin the presence or absence of an agonist such as adiol is optionallycompared to the activity of a reference compound such as ADEK or anothercompound disclosed herein in the same or a suitable similar assay. Suchinformation can then be used to characterize the test compound'scapacity to antagonize the activity of androgen receptor agonists suchas adiol. In these methods, test compounds can be assayed at two, three,four or more concentrations that range from about 0.01 nM to about 10mM, e.g., at one or more of about 0.01 nM, 0.1 nM, 1.0 nM, 10 nM, 100nM, 1 μM, 5 μM, 10 μM, 50 μM, 100 μM, 500 μM, 1 mM and 10 mM. Testcompounds that are capable of antagonizing Adiol-stimulated AR activitycan then be used to treat the conditions described herein.

Such assays can be performed essentially as described in the examplesdescribed herein, e.g., by contacting the test compound with a suitableAR assay system (under suitable conditions and for a sufficient time) inthe presence and/or absence of an AR agonist such as Adiol. Any of theseassays can optionally be performed in the presence or absence of otherAR modulators such as DHT, testosterone, HF or casodex to characterizethe effects of a test compound or a compound described herein to affectthe activity of such AR modulators. Other indirect assays, e.g.,measurement of PSA, can optionally also be used to characterize thecompounds.

The present invention also provides methods of inhibiting androgenreceptors in vitro or in vivo comprising contacting an androgen receptorwith an effective amount of a compound having the structure III, asdescribed above. In some embodiments of such methods, thetransactivation of androgen receptor is suppressed. In otherembodiments, the androgen receptor is mutant or native androgenreceptor.

As is apparent from the foreging, the invention provides method ofinhibiting an androgen receptor in vitro or in vivo comprisingcontacting the androgen receptor with an effective amount of a compounddisclosed herein, e.g., a compound having the structure I, II or III, ora prodrug of the compound, a pharmaceutically acceptable salt of thecompound, a stereoisomer of the compound, a tautomer of the compound, ora solvate of such compounds. In these methods, exemplary compounds ofstructure III, include compounds where the compound comprises a1,3-diene, 1,5-diene, 1,6-diene, 1,7-diene, 1,8-diene, 1,15-diene,1,16-diene, 3,16-diene, 4,8-diene, 1,3,5-triene, 1,4,6-triene,1,3,16-triene, 1,5,7-triene, 1,5,15-triene, 1,8,15-triene,1,5,16-triene, or 1,5,7,15-tetraene, within the fused four-ring system.In these methods, transactivation of androgen receptor can be detectablysuppressed for mutant or native androgen receptors. In some of theseembodiments, K is —CR¹R²—, e.g., —CH(OH)—, —C(CH₃)(OH)—, —C(CH₃)(ester)-or —C(C≡CH)(OH)— where the hydroxyl or ester is in the α- orβ-configuration.

In still other embodiments of methods of inhibiting androgen receptorsin vitro or in vivo, the compound comprises a 1,5-diene within the fusedfour-ring system. In other such embodiments, K is —CR¹R²—, R¹ is —OR¹¹,or R¹¹ is —H, substituted or unsubstituted alkyl, —C(O)—R¹²,—C(O)—NR¹²R¹³, or —C(O)—OR¹². In further embodiments, the compoundcomprises a 1,5-diene within the fused four-ring system, and R¹¹ is —H,or —C(O)—R¹². Thus, for example, the present invention provides methodsof inhibiting androgen receptors in vitro or in vivo, the compoundhaving the structure

Compounds of structure I, II, or III may be synthesized from knownstarting materials as shown in Schemes 1-17 and the Examples. By way ofexample and not limitation, 1,5-dienes such as compounds 3-6, may besynthesized from the 1,4-diene, 1, using standard synthetictransformations. For example, protection of 1 as the 17-ethylene ketal,followed by isomerization of the 1,4-diene to the 1,5 diene under basicconditions gives 3. The latter compound may be further transformed to 5by reduction and esterification. Compound 5 may be deprotected to givecompounds 6 and 7, or may undergo carbonyl addition reactions to givecompounds such as 8, as in Scheme 2. Scheme 3 illustrates the synthesisof 1,4-diene derivatives that may be used as starting materials for thesynthesis of invention compounds. The remaining schemes show syntheticroutes to various dienes, trienes, and tetraenes of the invention, andone skilled in the art will recognize that these routes may be readilymodified to produce the desired compounds of the invention. In somecases, it will be convenient to use starting compounds that contain 1 or2 R and/or R′ moieties or where a variable group such as E or G issubstituted. In other cases, moieties are added to the steroid moleculeat R R′, E and/or G after synthesis.

The instant invention also provides for pharmaceutical compositions orformulations which may be prepared by mixing one or more compoundsdisclosed herein such as ADEK or compounds of structures I, II, or III,prodrugs thereof, pharmaceutically acceptable salts thereof,stereoisomers thereof, tautomers thereof, or solvates thereof, with oneor more pharmaceutically acceptable carriers, excipients, binders,diluents, lubricants or the like, collectively “carriers”. Thesecompositions can be used to treat or ameliorate a variety of disordersmediated by androgen receptors. The compositions of the inventions maybe used to create formulations to prevent, treat or ameliorateconditions disclosed herein such as prostrate cancer, and in particularandrogen-independent prostrate cancer, as well as antiandrogen inducedwithdrawal syndrome. Such compositions can be in the form of, forexample, granules, powders, tablets, capsules, syrup, suppositories,injections, emulsions, elixirs, suspensions or solutions. The instantcompositions can be formulated for various routes of administration, forexample, by oral administration, by nasal administration, by rectaladministration, subcutaneous injection, intravenous injection,intramuscular injections, or intraperitoneal injection. The followingdosage forms are given by way of example and should not be construed aslimiting the instant invention.

For oral, buccal, and sublingual administration, powders, suspensions,granules, tablets, pills, capsules, gelcaps, and caplets are acceptableas solid, liquid or gel dosage forms. These can be prepared, forexample, by mixing one or more compounds of the instant invention, orpharmaceutically acceptable salts or tautomers thereof, with at leastone additive such as a starch or other additive. Suitable additives aresucrose, lactose, cellulose sugar, mannitol, maltitol, dextran, starch,agar, alginates, chitins, chitosans, pectins, tragacanth gum, gumarabic, gelatins, collagens, casein, albumin, synthetic orsemi-synthetic polymers or glycerides. Optionally, oral dosage forms cancontain other ingredients to aid in administration, such as an inactivediluent, or lubricants such as magnesium stearate, or preservatives suchas paraben or sorbic acid, or anti-oxidants such as ascorbic acid,tocopherol or cysteine, a disintegrating agent, binders, thickeners,buffers, sweeteners, flavoring agents or perfuming agents. Tablets andpills may be further treated with suitable coating materials known inthe art.

Liquid dosage forms for oral administration may be in the form ofpharmaceutically acceptable emulsions, syrups, elixirs, suspensions, andsolutions, which may contain an inactive diluent, such as water.Pharmaceutical formulations and medicaments may be prepared as liquidsuspensions or solutions using a sterile liquid, such as, but notlimited to, an oil, water, an alcohol, and combinations of these.Pharmaceutically suitable surfactants, suspending agents, emulsifyingagents, may be added for oral or parenteral administration. Parenteralformulations will typically be sterile and may optionally contain abacteriostat, e.g., EDTA or EGTA.

As noted above, suspensions may include oils. Such oils include, but arenot limited to, peanut oil, sesame oil, cottonseed oil, corn oil andolive oil. Suspension preparation may also contain esters of fatty acidssuch as ethyl oleate, isopropyl myristate, fatty acid glycerides andacetylated fatty acid glycerides. Suspension formulations may includealcohols, such as, but not limited to, ethanol, isopropyl alcohol,hexadecyl alcohol, glycerol and propylene glycol. Ethers, such as butnot limited to, poly(ethyleneglycol), petroleum hydrocarbons such asmineral oil and petrolatum; and water may also be used in suspensionformulations.

For nasal administration, the pharmaceutical formulations andmedicaments may be a spray or aerosol containing an appropriatesolvent(s) and optionally other compounds such as, but not limited to,stabilizers, antimicrobial agents, antioxidants, pH modifiers,surfactants, bioavailability modifiers and combinations of these. Apropellant for an aerosol formulation may include compressed air,nitrogen, carbon dioxide, or a hydrocarbon based low boiling solvent.

Injectable dosage forms generally include aqueous suspensions or oilsuspensions which may be prepared using a suitable dispersant or wettingagent and a suspending agent. Injectable forms may be in solution phaseor in the form of a suspension, which is prepared with a solvent ordiluent. Acceptable solvents or vehicles include sterilized water,Ringer's solution, or an isotonic aqueous saline solution.Alternatively, sterile oils may be employed as solvents or suspendingagents. Preferably, the oil or fatty acid is non-volatile, includingnatural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.

For injection, the pharmaceutical formulation and/or medicament may be apowder suitable for reconstitution with an appropriate solution asdescribed above. Examples of these include, but are not limited to,freeze dried, rotary dried or spray dried powders, amorphous powders,granules, precipitates, or particulates. For injection, the formulationsmay optionally contain stabilizers, pH modifiers, surfactants,bioavailability modifiers and combinations of these.

For rectal administration, the pharmaceutical formulations andmedicaments may be in the form of a suppository, an ointment, an enema,a tablet or a cream for release of compound in the intestines, sigmoidflexure and/or rectum. Rectal suppositories are prepared by mixing oneor more compounds of the instant invention, or pharmaceuticallyacceptable salts or tautomers of the compound, with acceptable vehicles,for example, cocoa butter or polyethylene glycol, which is present in asolid phase at normal storing temperatures, and present in a liquidphase at those temperatures suitable to release a drug inside the body,such as in the rectum. Oils may also be employed in the preparation offormulations of the soft gelatin type and suppositories. Water, saline,aqueous dextrose and related sugar solutions, and glycerols may beemployed in the preparation of suspension formulations which may alsocontain suspending agents such as pectins, carbomers, methyl cellulose,hydroxypropyl cellulose or carboxymethyl cellulose, as well as buffersand/or preservatives.

Besides those representative dosage forms described above,pharmaceutically acceptable excipients and carriers are generally knownto those skilled in the art and are thus included in the instantinvention. Such excipients and carriers are described, for example, in“Remingtons Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991),which is incorporated herein by reference.

The formulations of the invention may be designed to be short-acting,fast-releasing, long-acting, and sustained-releasing as described below.Thus, the pharmaceutical formulations may also be formulated forcontrolled release or for slow release.

The instant compositions may also comprise, for example, micelles orliposomes, or some other encapsulated form, or may be administered in anextended release form to provide a prolonged storage and/or deliveryeffect. Therefore, the pharmaceutical formulations and medicaments maybe compressed into pellets or cylinders and implanted intramuscularly orsubcutaneously as depot injections or as implants such as stents. Suchimplants may employ known inert materials such as silicones andbiodegradable polymers.

Specific dosages may be adjusted depending on conditions of disease, theage, body weight, general health conditions, sex, and diet of thesubject, dose intervals, administration routes, excretion rate, andcombinations of drugs. Any of the above dosage forms containingeffective amounts are well within the bounds of routine experimentationand therefore, well within the scope of the instant invention.

A therapeutically effective amount of a compound of the presentinvention may vary depending upon the route of administration and dosageform. The preferred compound or compounds of the instant invention is aformulation that exhibits a high therapeutic index. The therapeuticindex is the dose ratio between toxic and therapeutic effects which canbe expressed as the ratio between LD₅₀ and ED₅₀. The LD₅₀ is the doselethal to 50% of the population and the ED₅₀ is the dose therapeuticallyeffective in 50% of the population. The LD₅₀ and ED₅₀ are determined bystandard pharmaceutical procedures in animal cell cultures orexperimental animals. In general, daily dosages of about 0.1 mg/kg toabout 400 mg/kg, typically about 0.5 mg/kg, about 1 mg/kg, about 4 mg/kgor about 6 mg/kg to about 10 mg/kg, about 20 mg/kg, about 40 mg/kg orabout 60 mg/kg can be effective for treating humans or other mammals.

Other embodiments include use of a compound of structure I, II or II asdescribed herein for the preparation of a medicament or for thepreparation of a medicament for the prevention, treatment oramelioration of a disease or condition or to slow the progression of adisease or condition as described herein.

All references cited herein are incorporated herein by reference intheir entireties.

The present invention, thus generally described, will be understood morereadily by reference to the following examples, which are provided byway of illustration and are not intended to be limiting of the presentinvention.

Example 1 Synthesis of 3β-acetoxy-17,17-ethylenedioxyandrosta-1,5-diene(ADEK) (5)

An exemplary synthesis method for the compound is shown below.

Step 1: Synthesis of 17,17-ethylenedioxyandrosta-1,4-dien-3-one (2). Toa solution of androsta-1,4-dien-3,17-dione (1, 10.0 g) in benzene (600ml) and ethylene glycol (90 ml) was added toluene-p-sulfonic acidmonohydrate (0.3 g), and the solution was refluxed for 8-10 hours with aDean-Stark apparatus for collecting water. The reaction mixture wascooled to room temperature, diluted with ethyl acetate and washedthoroughly with water, dilute sodium bicarbonate solution, water andfinally with brine. The organic phase was dried over anhydrous magnesiumsulfate, filtered and the organic solvent removed by rotary evaporator.The resultant solid was crystallized from methanol to give17,17-ethylenedioxyandrosta-1,4-dien-3-one (2) as a white crystallinesolid in 95% yield (11.0 g).

Step 2: Synthesis of 17,17-ethylenedioxyandrosta-1,5-dien-3-one (3). Toa solution of 17,17-ethylenedioxyandrosta-1,4-dien-3-one (2, 8.5 g) infreshly distilled (over calcium hydride) dimethyl sulfoxide (160 ml) wasadded finely powdered potassium-t-butoxide (5.0 g), and the solution wasstirred at 10° C. for 2 hours. The reaction mixture was poured intoice-water and extracted with ethyl acetate-diethyl ether (2:1 v/v).Water and the solvent employed were previously saturated with dry ice(carbon dioxide). The organic layer was washed with ice-water severaltimes and dried over anhydrous magnesium sulfate. Evaporation of thesolvent in vacuo below 35° C. gave 8.5 g of semi-crystalline solid.Recrystallization from methanol gave17,17-ethylenedioxyandrosta-1,4-dien-3-one (3), mp. 154-57° C. Theuncrystallized sample was used as such for the next step.

Step 3: Synthesis of 17,17-ethylenedioxyandrosta-1,5-dien-3β-ol (4). Thecrude semi-crystalline residue (3, 8.0 g) obtained above was dissolvedin methanol (500 ml), and to this solution was added sodium borohydride(5.0 g) in water (100 ml) under ice cooling and stirring. After stirringfor 1 hour at 0° C., the excess of sodium borohydride was decomposed byadding 400 ml of 50% aqueous acetone. After the solution stood at roomtemperature overnight, the deposited crystals were filtered, washedthoroughly with water and dried under vacuum. The crude product waspurified by column chromatography over silica gel (eluent: ethylacetate/petroleum ether, 1:3, v/v) and recrystallized with methanol toafford 17,17-ethylenedioxyandrosta-1,5-dien-3β-ol (4, 6.5 g). Mp.138-140° C.

Step 4: 3β-acetoxy-17,17-ethylenedioxyandrosta-1,5-diene (5). A mixtureof crude 17,17-ethylenedioxyandrosta-1,5-dien-3β-ol (4, 1.2 g) inpyridine (10 ml) and acetic anhydride (3 ml) was stirred at roomtemperature for 16 hours. The reaction mixture was poured into ice-waterand the compound was extracted with ether. The organic layer was washedwith ice-water several times followed by brine and dried over anhydrousmagnesium sulfate. Evaporation of the solvent in vacuo below 35° C. gave1.0 g of crude solid. Recrystallization from methanol gave3β-acetoxy-17,17-ethylenedioxyandrosta-1,5-diene (5). Mp. 105-6° C.

Example 2

An exemplary synthesis method for 3-hydroxyandrosta-1,5-dien-17-one (7)is shown below.

Step 1: 3β-acetoxyandrosta-1,5-dien-17-one (6). Ketal 5 (0.44 g) fromExample 1 was dissolved in acetone-water (30 ml, 8:2), and treated withtoluene-p-sulfonic acid monohydrate (0.1 g). After the mixture had beenstirred at room temperature for 16 hr it was concentrated to half of itsvolume and diluted with a cold half saturated sodium bicarbonatesolution. The solution was cooled, filtered to give white solid compound(0.38 g, 98%), which was further crystallized from methanol. M.p.185-87° C., purity 99% (LC-MS).

Step 2: 3β-hydroxyandrosta-1,5-dien-17-one (7). A mixture of compound 6(0.25 g) and potassium carbonate (0.3 g) in methanol-water (15 ml, 9:1)was stirred at room temperature for 8 hr. Solution was concentrateddiluted with cold water, cooled and the precipitated solid was filtered,and crystallized from methanol. White solid, mp. 138-40° C.

Example 3 Chemicals and Plasmids

DHT, Adiol, 17β-estradiol (E2), progesterone (P), and dexamethasone(Dex) were obtained from Sigma, Hydroxyflutamide (HF) was from Schering,and casodex was from ICI Pharmaceuticals. Other steroid compounds,derivatives of DHEA, were synthesized as above. pSG5-AR and pSG5-ARA70were obtained as in: Yeh, S., Miyamoto, H. & Chang, C. (1997) Lancet349, 852-853; Miyamoto, H., Yeh, S., Wilding, G. & Chang, C. (1998)Proc. Natl. Acad. Sci. USA 95, 7379-7384; Miyamoto, H., Yeh, S., Lardy,H., Messing, E. & Chang, C. (1998) Proc. Natl. Acad. Sci. USA 95,11083-11088; Chang, H.-C., Miyamoto, H., Marwah, P., Lardy, H., Yeh, S.,Huang, K.-E. & Chang, C. (1999) Proc. Natl. Acad. Sci. USA 96,11173-11177; Yeh, S. & Chang, C. (1996) Proc. Natl. Acad. Sci. USA 93,5517-5521.

Cell Culture, Transfection, and Reporter Gene Assay. The human prostatecancer cell lines, LNCaP, PC-3, and DU145, and non-prostate cancer cellline COS-1 were maintained in RPMI or Dulbecco's modified Eagle's medium(DMEM) (Life Technologies) supplemented with 10% fetal bovine serum(FBS). Transfections and luciferase (Luc) assays were performed aspreviously described (Miyamoto, H., Yeh, S., Wilding, G. & Chang, C.(1998) Proc. Natl. Acad. Sci. USA 95, 7379-7384; Miyamoto, H., Rahman,M., Takatera, H., Kang, H.-Y., Yeh, S., Chang, H.-C., Nishimura, K.,Fujimoto, N. & Chang, C. (2002) J. Biol. Chem. 277, 4609-4617). Briefly,cells seeded to reach a density of 50-60% confluence in 12 well tissueculture plates were transfected with 1.5 μg of DNA according to“SuperFect transfection” instructions (Qiagen). After 2-3 h incubation,cells were treated with medium supplemented with charcoal-stripped FBScontaining either ethanol or ligands for 24 h. The cells were thenharvested and whole cell extracts were used for Luc assay. The Lucactivity was determined using a Dual-Luciferase Reporter Assay System(Promega) and luminometer.

Western Blot. Western blotting analysis was performed in LNCaP cells,using monoclonal PSA antibody (DAKO), as described previously (Miyamoto,H., Rahman, M., Takatera, H., Kang, H.-Y., Yeh, S., Chang, H.-C.,Nishimura, K., Fujimoto, N. & Chang, C. (2002) J. Biol. Chem. 277,4609-4617). An antibody for -actin (Santa Cruz Biotechnology) was usedas the internal control. Blots were quantitated by Collage software(Fotodyne).

Ligand Binding Assay. Whole cell extracts from COS-1 with transienttransfection of pSG5-AR, or LNCaP without transfection, were incubatedfor 2 h at 37° C. with 1 nM [³H]-synthetic androgen methyltrienolone(R1881) in the presence and absence of increasing concentrations(1-10,000 nM) of unlabeled ligands. Then, hydroxyapatite (Bio-Rad) wasadded and stirred for 15 min at 4° C. After centrifugation and washing,radioactivity was determined by scintillation counting.

Anti-DHT Effect of DHEA Derivatives with Low Androgenic Activity on ARTranscription. Test compounds as antiandrogenic compounds, werecharacterized by analysis of their ability to induce AR transcriptionalactivity in the AR-negative PC-3 cell line. The Luc activity wasdetermined in the cell extracts with transient transfection of wild-typeAR plasmid and androgen response element-reporter plasmid (mouse mammarytumor virus (MMTV)-Luc). After transfection, the cells were treated withvarious compounds at 0.1-1,000 nM. Of 17 compounds tested, only four(No. 5: 3β,7α,17β-trihydroxyandrost-5-ene; No. 10: ADEK; No. 14:3β-acetoxyandrost-1,5-diene-17-one; and No. 16:3β-hydroxyandrost-1,5-diene-17-one) at 1000 nM showed marginal inductionon AR transcription, as compared to mock treatment.

Compounds 5, 10, 14 and 16 were characterized for their anti-DHTactivity on AR transcription in PC-3 cells. Cells were transfected withAR plasmid and MMTV-Luc reporter in the presence of 1 nM DHT and each ofthese compounds at 0.01, 0.1, or 1 μM. While compounds No. 5, No. 14,and No. 16 showed modest suppression on DHT-induced AR transactivation,ADEK suppressed it to 30% in a dose-dependent manner. Some compounds,3β-acetoxy-17β-hydroxyandrost-1,5-diene,7α-hydroxyandrost-5-ene-3,17-bis ethylene ketal,7β,17β-dihydroxyandrost-5-ene-3-ethylene ketal,3β,16α-bis-carbomethoxyandrost-5-ene-7,17-dione,3β,17β-dihydroxyandrost-4-ene and androst-1,4-diene-3,17-dione, wereless effective in inhibiting DHT-induced AR transactivation. Othercompounds, 3β,7α,17β-trihydroxyandrostane, 16α-bromoepiandrosterone,3β,7β,17β-trihydroxyandrost-5-ene, 3β-hydroxy-5α-androstane-17-one,7α-hydroxyandrost-5-ene-3,17-bis ethylene ketal,17β-acetoxyandrost-4-ene-3,6-dione and17β-propionony-7-oxoandrost-5-en-3-ethylene ketal, had little or nocapacity to inhibit DHT-induced AR transactivation.

To accomplish these studies, PC-3 cells were transfected with thewild-type AR expression plasmid pSG5-AR and MMTV-Luc. Aftertransfection, cells were cultured for 24 h with 1 nM DHT or 1,000 nM ofvarious DHEA derivatives. The Luc activity is presented relative to thatin the presence of DHT (set as 100%). The Luc activity was measuredrelative to that of ethanol treatment (set as 1-fold). Values from themean±SD of at least three determinations were used. PC-3 cells weretransfected with the pSG5-AR and MMTV-Luc. After transfection, cellswere cultured for 24 h with various concentrations of compounds No. 5,10 (ADEK), 14, or 16 in the presence of 1 nM DHT. The Luc activity wasdetermined relative to that in the presence of DHT (set as 100%). Valuesfrom the mean±SD of at least three determinations were obtained.

ADEK was further investigated, using different cell lines and differentreporters, and was also compared to non-steroidal antiandrogens, HF andcasodex. ADEK had lower androgenic activity on wild-type ARtranscription than HF and casodex in COS-1 cells. ADEK at 1 μMsuppresses DHT-induced wild-type AR transcription to 21%, similar to thesuppression by HF and casodex. In LNCaP cell line, 10 μM HF acts as fullagonist, and therefore shows no suppression of DHT-induced mutant ARtranscription, consistent with the previous findings (Kuil, C. W. &Mulder E. (1996) Endocrinology 137, 1870-1877; Miyamoto, H. & Chang, C.(2000) Int. J. Urol. 7, 32-34). Casodex and ADEK exhibiteddose-dependent suppression to 22% and 17%, respectively, and androgenicactivity of ADEK was lower than that of casodex. Similar results wereobtained when MMTV-Luc was replaced with PSA-Luc. In addition, one ofthe AR coactivators, ARA70, which has been shown to enhancesignificantly agonist activity of antiandrogens (5-12 fold) (Yeh, S.,Miyamoto, H. & Chang, C. (1997) Lancet 349, 852-853; Miyamoto, H., Yeh,S., Wilding, G. & Chang, C. (1998) Proc. Natl. Acad. Sci. USA 95,7379-7384), marginally enhanced AR transactivation in the presence ofADEK (<2-fold) in DU145 cells. The results indicated that ADEK acts as apotent antagonist on DHT-enhanced transactivation of both wild-type ARand a mutant AR. Several compounds related to ADEK, i.e.,3β-acetoxyandrosta-1,5-dien-17-one, androsta-1,4-dien-3,17-dione,3β-hydroxyanrosta-1,5-dien-17-one and3β-acetoxy-17β-hydroxyandrosta-1,5-dien, did not show significantantagonistic effects. The agonist effect of ADEK was marginal and lowerthan that of non-steroidal antiandrogens. Because of this, there is lesspossibility of inducing withdrawal response in prostate cancer patientswhen using compounds such as ADEK.

The effects of ADEK on the DHT-induced transcriptional activity of ARwas examined in COS-1 or LNCaP cells transfected with MMTV-Luc. ThepSG5-AR was co-transfected in COS-1 cells. After transfection, cellswere cultured for 24 h in the presence or absence of 1 nM DHT or variousconcentrations of HF, casodex, or ADEK. Luc activity was analyzedrelative to Luc activity in the presence of DHT (set as 100%). Valueswere obtained from the mean±SD of at least three determinations. DU145cells were transfected with the pSG5-AR and MMTV-Luc in the presence orabsence of pSG5-ARA70. After transfection, cells were cultured for 24 hwith various concentrations of HF, casodex, or ADEK. The Luc activity ispresented relative to that of ETOH treatment without ARA70 (set as1-fold). Values were obtained for the mean±SD of at least threedeterminations. For COS-1 cells, relative Luc activity in 1 μM ADEK was21% of the level relative to the DHT control (100%), while Luc activityin 0.1 μM ADEK and 0.01 μM respectively was about 60% and about 75% ofthe control level.

Example 4 Anti-DHT Effect of ADEK on PSA Expression and CellProliferation

The PSA is an AR responsive gene and presently the most useful tumormarker to monitor prostate cancer progression. The capacity of ADEK tomodulate PSA expression in prostate cancer cells was tested. The Westernblotting assay showed that DHT increased endogenous PSA expression inLNCaP cells to 4.3-fold over mock treatment and that ADEK and casodexdecreased DHT-induced PSA expression to 49% and 58%, respectively. HFinduces PSA expression to 3.5-fold, whereas ADEK and casodex increase itto less than 2-fold. The effect of ADEK on cell growth of LNCaP wastested. DHT significantly increased cell growth, and ADEK and casodexantagonized this DHT effect. ADEK and casodex marginally increasedgrowth in the absence of androgen. These results confirm the ARtranscription data and suggest that ADEK can inhibitandrogen-AR-mediated prostate cancer progression.

To accomplish the PSA expression analysis, cell extracts from LNCaPcells cultured for 48 h with 1 μM HF, 1 μM casodex, or 1 μM ADEK in thepresence or absence of 1 nM DHT, were analyzed on Western blots using anantibody to the PSA. The 33-kDa protein was quantitated. β-Actinexpression was used as an internal control. The normalized expressionlevel in the DHT treated cells was set as 100%. The mean±SD of threeseparate experiments was determined. To determine the effect on LNCaPcell growth, the cells were cultured with 1 μM HF, 1 μM casodex, or 1 μMADEK in the presence or absence of 1 nM DHT. Total cell number wascounted by hemocytometer. The mean of at least three determinations wasdetermined.

Example 5 Interruption of Androgen Binding to the AR by ADEK

Clinically available antiandrogens have an affinity for the AR, allowinga competition with androgens for binding. To determine whether ADEK hasthis common feature of AR antagonists, the competitive androgen bindingassay was performed. The affinity of ligands for the AR was assessed byincubating whole cell extracts of LNCaP or COS-1 with transfectedwild-type AR with 1 nM [³H]-R1881 in the presence of variousconcentrations (1-10,000 nM) of unlabeled DHT, HF, casodex, or ADEK. Asdescribed previously (Schuurmans, A. L. G., et al. (1988) Int. J. Cancer42, 917-922), the relative binding affinity (RBA) values were calculatedfrom the constructed competitive binding curves as the ratio ofconcentration of unlabeled ligand and concentration of DHT required toinhibit [³H]-R1881 binding by 50% (Table 1 below). Competitive RBAs inLNCaP cells were DHT>casodex>HF>ADEK. Similar results were obtained inwild-type AR transfected COS-1 cells, although the RBAs are lower andbinding of all the compounds in competition with [³H]-R1881 was weaker.These results confirm that ADEK also competes significantly withandrogen for AR binding.

TABLE 1 Ligand RBA in LNCaP RBA in COS-1 with AR DHT 100.0 100.0 HF 23.017.1 Casodex 36.4 25.5 ADEK 11.1 6.0

Example 6 Anti-Adiol Effect of ADEK on AR Transcription

Adiol, which is produced from DHEA and which can be converted totestosterone, possesses intrinsic androgen activity. Among androgens itis unique in that both HF and casodex failed to block significantlyAdiol-induced AR transactivation in prostate cancer cells. Becausecastration with or without combination therapy with antiandrogen,decreases the serum concentration of Adiol by only 40-50% (Bélanger, etal. (1986) J. Clin. Endocrinol. Metab. 62, 812-815; Labrie, F., et al.(1988) Br. J. Urol. 61, 341-346), previous findings suggested thatcurrent CAB treatment might be insufficient to block Adiol's action inAR-positive prostate cancer. Therefore, the capacity of ADEK to inhibitAdiol-induced AR transcription was analyzed by measuring MMTV-Lucactivity. The wild-type AR expression plasmid pSG5-AR was co-transfectedin PC-3 cells to permit assay of Adiol-induced AR transcription. Aftertransfection, cells were cultured for 24 h in the presence or absence of2.5 nM Adiol and 1 μM HF, 1 μM casodex, or 1 μM ADEK. Adiol at 2.5 nMincreased AR transcriptional activity in PC-3 and LNCaP to 4.5-fold and2.8-fold, respectively, over mock treatment. ADEK at 1 μM repressedAdiol-induced AR transcription by about 43% and 58% in PC-3 and LNCaP,respectively, whereas HF at 1 μM and casodex at 1 μM failed tosignificantly block Adiol-induced AR transcription. These resultssupport the conclusion that ADEK can suppress AR transactivation inducedby classic androgens as well as by adrenal androgen. In these assays,Luc activity was presented relative to that in the presence of Adiol(set as 100%). Values were obtained from the mean±SD of at least threedeterminations. Other test compounds, e.g., compounds of structure I, IIor III, can be characterized for their capacity to modulate orantagonize adiol-induced AR transcription, or transcription induced byother AR modulators, in essentially the same manner using this assay ora suitable variation of this assay, e.g., use of adiol at otherconcentrations such as 1, 2, 3, 4 or 5 nM.

Example 7 Steroid Hormone Specificity of ADEK

To characterize the steroid hormone activity of ADEK, PC-3 cells weretransfected with steroid receptor/reporter (progesterone receptor(PR)/MMTV-Luc, glucocorticoid receptor (GR)/MMTV-Luc, or estrogenreceptor (ER)/ERE-Luc) and analyzed for expression of the Luc reportergene. After transfection, the cells were cultured for 24 h in thepresence or absence of ligand (10 nM DHT, 10 nM progesterone, 10 nMdexamethasone, or 10 nM 17β-estradiol) or ADEK at 0.01 mM, 0.1 mM and1.0 mM. The Luc activity was measured relative to that of ETOH treatment(set as 1-fold). Values were obtained from the mean±SD of at least threedeterminations. The results indicated that ADEK had some estrogenicactivity, but ADEK had no significant progesterone, glucocorticoidactivity or androgenic activity.

1. A compound having the structure

or a pharmaceutically acceptable salt of the compound wherein, A is—CR⁹R¹⁰; E is —C(O)— or —CR⁵R⁶—; G is ═CR³—; K is —CR¹R²—; R and R′independently are —F, —Cl, —Br, —I, substituted or unsubstituted loweralkyl, substituted or unsubstituted lower alkene, substituted orunsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted or unsubstituted lower alkoxy, and—S(O)₀₋₂R¹⁴; R¹ is —OR¹¹, —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹²,—C(S)—R¹², —C(S)—OR¹² or —S(O)₀₋₂—R¹²; R² is —H, substituted orunsubstituted lower alkyl, substituted or unsubstituted lower alkene orsubstituted or unsubstituted lower alkyne; R³ and R⁵ independently are—H, —F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl,substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted lower alkoxy or—S(O)₀₋₂R¹⁴; R⁶ is —H, —F, —Cl, —Br, —I, substituted or unsubstitutedlower alkyl, substituted or unsubstituted lower alkene, substituted orunsubstituted lower alkyne, —CN, —COR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—OC(O)—R¹⁵, —OH, substituted and unsubstituted lower alkoxy or and—S(O)₀₋₂R¹⁴; R⁷ and R⁸ independently are —H or substituted orunsubstituted lower alkyl; R⁹ is —OH or substituted or unsubstitutedlower alkoxy; R¹⁰ is substituted or unsubstituted lower alkyl,substituted or unsubstituted lower alkene, substituted or unsubstitutedlower alkyne, substituted or unsubstituted —S(O)₀₋₂(lower alkyl), or R⁹and R¹⁰, together with the carbon to which they are attached, form a 5-,6-, or 7-member heterocyclyl or cycloalkyl group; R¹¹ is —H, substitutedor unsubstituted alkyl, substituted or unsubstituted alkene, substitutedor unsubstituted alkyne, —C(O)—R¹², —C(O)—NR¹²R¹³, C(O)—R¹², —C(S)—R¹²,—NR¹²R¹³, —S(O)₂—R¹², —S(O)₂—OR¹², and —P(O)(OR¹²)(OR¹³)₀₋₁; R¹² and R¹³independently are —H, substituted or unsubstituted alkyl, substituted orunsubstituted alkene, substituted or unsubstituted alkyne, substitutedor unsubstituted aryl, substituted or unsubstituted arylalkyl,substituted or unsubstituted heterocyclyl or substituted orunsubstituted heterocyclylalkyl; R¹⁴ and R¹⁵ independently are —H,substituted or unsubstituted lower alkyl, substituted or unsubstitutedlower alkene, substituted or unsubstituted lower alkyne, substituted orunsubstituted C₆₋₁₀ aryl, or substituted or unsubstituted C₇₋₁₂arylalkyl; n is 0, 1, or 2; and n′ is 0, 1 or
 2. 2. The compound ofclaim 1 wherein R¹ is —OR¹¹, —C(O)—R¹² or —C(O)—OR¹².
 3. The compound ofclaim 1 wherein E is —CR⁵R⁶—.
 4. The compound of claim 3 wherein R⁵ is—OH.
 5. The compound of claim 1 having the structure


6. The compound of claim 5 having the structure

wherein R¹⁶ is H, alkyl or fluoroalkyl.
 7. The compound of claim 6wherein R¹⁶ is H.
 8. The compound of claim 5 having the structure


9. The compound of claim 1 having the structure


10. The compound of claim 9 having the structure

wherein R¹⁶ is H, alkyl or fluoroalkyl.
 11. The compound of claim 10wherein R¹⁶ is —H.
 12. The compound of claim 9 having the structure


13. A pharmaceutical formulation comprising one or more excipients and acompound having the structure

or a pharmaceutically acceptable salt of the compound wherein, A is—CR⁹R¹⁰; E is —C(O)— or —CR⁵R⁶—; G is ═CR³—; K is —CR¹R²—; R and R′independently are —F, —Cl, —Br, —I, substituted or unsubstituted loweralkyl, substituted or unsubstituted lower alkene, substituted orunsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted or unsubstituted lower alkoxy, and—S(O)₀₋₂R¹⁴; R¹ is —OR¹¹, —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹²,—C(S)—R¹², —C(S)—OR¹² or —S(O)₀₋₂—R¹²; R² is —H, substituted orunsubstituted lower alkyl, substituted or unsubstituted lower alkene orsubstituted or unsubstituted lower alkyne; R³ and R⁵ independently are—H, —F, —Cl, —Br, —I, substituted and unsubstituted lower alkyl,substituted and unsubstituted lower alkene, substituted andunsubstituted lower alkyne, —CN, —COOR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted lower alkoxy or—S(O)₀₋₂R¹⁴; R⁶ is —H, —F, —Cl, —Br, —I, substituted or unsubstitutedlower alkyl, substituted or unsubstituted lower alkene, substituted orunsubstituted lower alkyne, —CN, —COR¹⁴, —C(O)NR¹⁴R¹⁵, —NO₂, —NR¹⁴R¹⁵,—NR¹⁴—C(O)—R¹⁵, —OH, substituted and unsubstituted lower alkoxy or and—S(O)₀₋₂R¹⁴; R⁷ and R⁸ independently are —H or substituted orunsubstituted lower alkyl; R⁹ is —OH or substituted or unsubstitutedlower alkoxy; R¹⁰ is substituted or unsubstituted lower alkyl,substituted or unsubstituted lower alkene, substituted or unsubstitutedlower alkyne, substituted or unsubstituted —S(O)₀₋₂(lower alkyl), or R⁹and R¹⁰, together with the carbon to which they are attached, form a 5-,6-, or 7-member heterocyclyl or cycloalkyl group; R¹¹ is —H, substitutedor unsubstituted alkyl, substituted or unsubstituted alkene, substitutedor unsubstituted alkyne, —C(O)—R¹², —C(O)—NR¹²R¹³, —C(O)—OR¹²,—C(S)—R¹², NR¹²R¹³, —S(O)₂—R¹², —S(O)₂—OR¹², and —P(O)(OR¹²)(OR¹³)₀₋₁;R¹² and R¹³ independently are —H, substituted or unsubstituted alkyl,substituted or unsubstituted alkene, substituted or unsubstitutedalkyne, substituted or unsubstituted aryl, substituted or unsubstitutedarylalkyl, substituted or unsubstituted heterocyclyl or substituted orunsubstituted heterocyclylalkyl; R¹⁴ and R¹⁵ independently are —H,substituted or unsubstituted lower alkyl, substituted or unsubstitutedlower alkene, substituted or unsubstituted lower alkyne, substituted orunsubstituted C₆₋₁₀ aryl, or substituted or unsubstituted C₇₋₁₂arylalkyl; n is 0, 1, or 2; and n′ is 0, 1 or
 2. 14. The pharmaceuticalformulation of claim 13 wherein R⁷ and R⁸ are —CH₃.
 15. Thepharmaceutical formulation of claim 14 wherein the compound has thestructure

wherein R¹⁶ is H, alkyl or fluoroalkyl.
 16. The pharmaceuticalformulation of claim 15 wherein R¹⁶ is —H.
 17. The pharmaceuticalformulation of claim 14 wherein the compound has the structure


18. The pharmaceutical formulation of claim 14 wherein the compound hasthe structure

wherein R¹⁶ is H, alkyl or fluoroalkyl.
 19. The pharmaceuticalformulation of claim 18 wherein R¹⁶ is —H.
 20. The pharmaceuticalformulation of claim 14 wherein the compound has the structure